LOCUS       sequence-409840-        7669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ddGFP-B3-inserted Pcdh-alpha-4 lacking an intracellular domain 
            (ICD).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7669)
  AUTHORS   Kanadome T, Hoshino N, Nagai T, Yagi T, Matsuda T
  TITLE     Visualization of trans-interactions of a protocadherin-alpha between
            processes originating from single neurons.
  JOURNAL   iScience. 2023 Jul 17;26(7):107238. doi: 10.1016/j.isci.2023.107238.
            eCollection 2023 Jul 21.
  PUBMED    37534169
REFERENCE   2  (bases 1 to 7669)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "iScience.";
            date: "2023-07-17"; volume: "26(7):107238. doi"; pages: " 
            10.1016/j.isci.2023.107238. eCollection 2023 Jul 21"
COMMENT     Sequence Label: pCX:B3-alpha-4-Delta-ICD
FEATURES             Location/Qualifiers
     source          1..7669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1733..1742
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4658..4684
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4691..4710)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4756..4811
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4810..4829)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5172..5188)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5172..5188)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5185..5207)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5196..5212
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5220..5226,5227..5244,5245..5250))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5265..5286
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5345..5540
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5391..5526
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5546..5680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5583..5602)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5637..5656
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5748..5765)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5919..6507)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5999..6018)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6678..7469,7470..7538))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7301..7320
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7539..7643)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggsgc gccggggggg gggggggggg gggggggggg
      541 ggggggggsg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gkgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg rgggctcggg ggrggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcgccaccat
     1741 ggaattttcc tggggaagtg gccaggaatc ccagcgcttg cttctttctt ttctgcttct
     1801 tgcaatctgg gaggcaggga acagcgctag cgcaaccatc aaagaattta tgaggttcaa
     1861 agtacgcatg gagggtagta tgaacgggca tgagttcgaa attgaaggag aaggggaagg
     1921 ccgcccctat gaggggaccc aaaccgccaa acttaaagtg acaaagggag gtccacttcc
     1981 ttttgcctgg gatatattga gcccacagtt catgtacgga agcgaagcct atgtacgaca
     2041 tcctgcagac ataccagatt acaagaagct gccctttcca gaaggcttta agtgggaaag
     2101 agtgatgaac tttgaagatg gcggcttggt aacagtgact caggacagca gtctccaaga
     2161 tggaactttg atctgtaagg tcaaaatgag gggtactaac tttccccccg atgggcccgt
     2221 catgcaaaaa aagacaatgg gctgggaagc ctcaaccgag atgttgtacc cagaagatgg
     2281 agtgctcaag ggacattctt atcaggcatt gaagctgaaa gatggcggac attacttggt
     2341 tgagtttgag actatctata tggccaagaa gcccgtacaa cttcctggag attactgcgt
     2401 agacaccaaa ctggatataa cctctcataa cgaggattac accattgtcg agcagtacgg
     2461 ccgcagtgaa ggtagacata gacttggaat ggatgagcga tataaggcta gccagatcca
     2521 ctactccatc cctgaggagg ccaaacacgg caccttcgtg ggccgcatcg cgcaggacct
     2581 ggggctggag ctgacggagc tggtgccccg cctgttcaga gtggcgtcca aggaccgcgg
     2641 agaccttctg gaggtaaatc tgcagaatgg cattttgttt gtgaactctc ggatcgaccg
     2701 ggaggagctg tgcgggcgga gcgcggagtg cagcatccac ctggaggtga tcgtggacag
     2761 gccgttgcag gttttccacg tggaggtgga ggtgagggac attaacgaca accctcccag
     2821 gttcccaaca acacaaaaga atctgttcat tgcagaatca aggccacttg acacttggtt
     2881 tccactagag ggcgcttcag acgcagatat cggaatcaat gctgtactga cttacagact
     2941 gagtccaaat gattactttt ctttggaaaa accatccaac gacgaacggg taaaaggtct
     3001 tggacttgta ttacggaaat ctttagaccg ggaggaaact ccagagataa ttttagtgct
     3061 tactgtcacg gacggaggaa agccagagct gaccggcagt gttcagttac tcatcactgt
     3121 gctggatgcc aatgataatg ctccagtttt tgacagatcc ctgtataccg tgaaattacc
     3181 agaaaacgtt ccaaatggga cattggtagt caaagtcaat gcctcagatt tagacgaagg
     3241 ggtaaatggg gatattatgt actcattttc tacagatatt tcaccaaatg tgaaatacaa
     3301 attccacata gaccctgtta gcggagagat tattgtaaag ggatacattg attttgaaga
     3361 atgcaaatcc tatgaaattc tcatagaggg aattgacaag ggacaacttc cactctctgg
     3421 gcactgtaaa gtcattgtac aagttgaaga catcaacgat aatgttccag aattggaatt
     3481 taaatctcta tcacttccaa tacgagagaa ctctccagtg ggcactgtca tcgcactcat
     3541 tagtgtgtct gatcgggaca cgggtgtcaa cgggcaggtg acctgctccc tgacaagtca
     3601 tgtccccttc aagttggtgt ccacattcaa gaattactat tcgctcgtgc tggacagcgc
     3661 cctggacaga gagacaacag cggactataa ggtggtggtg acagcgcggg atgggggctc
     3721 tccctcgctg tgggccacgg ctagtgtgtc tgttgaggtt gctgacgtga acgacaatgc
     3781 acctgtgttc gcgcagcccg aatacacggt gttcgtgaag gagaacaacc cgcctggtgc
     3841 gcacatcttc acggtgtcag cgatggatgc ggacgcacag gagaacgcgc tggtgtccta
     3901 ctcgctggtg gagcggaggg tgggcgagcg cttgctgtcg agctatgtgt ctgtgcacgc
     3961 ggagagcggc aaggtgttcg cgctgcagcc tctggaccat gaggagctgg agctgctgcg
     4021 gttccaggtg agcgcgcggg atgctggtgt acctgccctg ggcagcaatg tgactctgca
     4081 ggtgtttgtg ctggacgaga atgacaacgc gcccacactg ctggaacctg aggcaggagt
     4141 ctctggtgga atcgtgagcc ggttggtgtc cagatcagtg ggtgcaggcc atgtggtggc
     4201 taaggtgcgc gcggtggatg cagactctgg ctataatgca tggctctctt atgagctgca
     4261 atcgtcagaa ggcaattccc gtagcctttt ccgcgtaggt ttgtatacgg gcgagattag
     4321 tactacgcgc atactggatg aagcagattc gccacgtcag cgccttctgg tgctggtgaa
     4381 ggaccatggt gacccagcaa tgattgttac cgccacagtg ttggtgtctc tggtagagaa
     4441 tggcccggta ccaaaggctc catcgcgagt atccacgagt gtcacacact ctgaggcgtc
     4501 actggtggat gtcaacgtgt acctgatcat tgccatctgt gcagtgtcca gcctgctagt
     4561 gctcacgctg ctgctgtaca cagcgctgcg ctgttccact gtccccagtg agagcgtgtg
     4621 cgggcctcca aaaccggtaa tggtgtgctc cagtgcatat ccgtacgacg taccggatta
     4681 tgcgtgaaga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
     4741 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
     4801 tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt
     4861 atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc
     4921 tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa
     4981 aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa
     5041 catccctaaa attttcctta catgttttac tagccagatt tttcctcctc tcctgactac
     5101 tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag cccaagcttg
     5161 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5221 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5281 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg
     5341 atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     5401 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5461 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5521 gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata
     5581 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5641 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc
     5701 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5761 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5821 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5881 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5941 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6001 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6061 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6121 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6181 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6241 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6301 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6361 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6421 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6481 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6541 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6601 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6661 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6721 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6781 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6841 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6901 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6961 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7021 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7081 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7141 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7201 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7261 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7321 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7381 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7441 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7501 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7561 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7621 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctg
//
